CN113475746B - Secondary extraction method of tobacco extract residues - Google Patents
Secondary extraction method of tobacco extract residues Download PDFInfo
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- CN113475746B CN113475746B CN202110687892.7A CN202110687892A CN113475746B CN 113475746 B CN113475746 B CN 113475746B CN 202110687892 A CN202110687892 A CN 202110687892A CN 113475746 B CN113475746 B CN 113475746B
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/20—Biochemical treatment
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- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/26—Use of organic solvents for extraction
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Abstract
The invention discloses a secondary extraction method of tobacco extract residues, which comprises the following steps: inoculating the colony of micrococcus ZY02 with the preservation number of CGMCC NO.21313 into a liquid culture medium to prepare a seed solution; mixing and fermenting the micrococcus seeds and the residue mixed solution to obtain a fermentation residue mixed solution; filtering the mixture of the fermentation residues to obtain a first filtrate and a first solid, and centrifuging the first filtrate to obtain a second solid; shaking a third solid obtained by mixing the first solid and the second solid with absolute ethyl alcohol, and filtering to obtain a second filtrate; mixing the supernatant with the second filtrate to obtain a secondary tobacco extract, and vacuum-concentrating the secondary tobacco extract to obtain a concentrated solution; carrying out reflux extraction treatment on the concentrated solution to obtain a baked concentrated solution; adding absolute ethyl alcohol into the baking concentrated solution, and centrifuging to obtain an alcohol precipitate; adding 1, 2-propylene glycol into the alcohol precipitate. The secondary extraction method of the tobacco extract residues disclosed by the invention can effectively extract beneficial components in the tobacco extract residues to obtain the novel tobacco extract.
Description
Technical Field
The invention relates to the field of tobacco waste utilization, in particular to a secondary extraction method of tobacco extract residues.
Background
The tobacco extract is an extremely important tobacco flavor in cigarette processing, and can endow the cigarette with more natural and full aroma.
The tobacco extract is generally prepared by pulverizing tobacco leaves or pieces into tobacco powder, and extracting, purifying and concentrating the flavor components in the tobacco powder by using organic substances or water as a solvent. During the extraction process, the components which are easy to dissolve in the solvent are extracted, and some solid matters such as fiber, protein, starch, pectin, lignin, a part of flavor components wrapped inside are difficult to extract, so that the residue of the tobacco powder is formed.
These tobacco residue is typically discarded, causing waste. And certain high-quality tobacco resources have higher prices, and if residues left after the residues are used for preparing the tobacco extract can be extracted for the second time to prepare a new tobacco extract, better economic and social benefits can be generated.
The tobacco extract residue is subjected to secondary extraction, firstly, macromolecular substances are degraded, on one hand, the macromolecular substances are converted into micromolecules and flavor components, and on the other hand, the flavor components wrapped in the tobacco are better released. The macromolecular substances in the residue can be degraded by chemical and biological methods. The chemical method mainly comprises a strong acid or strong alkali method, and the method is difficult to solve the residual acid and alkali and is not suitable for preparing the tobacco extract. The biological method mainly degrades and converts macromolecular substances into soluble sugar, amino acid and other small molecules through microbial or enzyme preparation treatment, and further can induce Maillard reaction under high temperature condition to generate more fragrance components. Some aroma-enhancing microorganisms can also utilize sugar and amino acid to generate aroma substances, and the tobacco extract residues are treated by a biological method and subjected to secondary extraction, so that the method has better practical value.
Therefore, how to provide a method for treating the tobacco extract residue by a biological method which can effectively extract beneficial components in the tobacco extract residue becomes a technical problem which needs to be solved in the field.
Disclosure of Invention
An object of the present invention is to provide a new technical solution for a biological process for treating tobacco extract residue, which can effectively extract beneficial components in the tobacco extract residue.
According to a first aspect of the present invention, a method for secondary extraction of tobacco extract residue is provided.
The secondary extraction method of the tobacco extract residue comprises the following steps:
step (1): inoculating a colony of Micrococcus (Micrococcus Pribram) ZY02 with the preservation number of CGMCC NO.21313 into a liquid culture medium, and culturing for 6-36h in a shaker at 25-40 ℃ and 100-200r/min to prepare a seed solution;
step (2): centrifuging the seed solution at 4 deg.C at 4000r/min for 20min, removing supernatant, adding sterile water for redissolving, and adjusting OD 600 2.0, preparing micrococcus seeds;
and (3): mixing the tobacco extract residue with deionized water according to the mass ratio of 1 (1-10) to obtain a residue miscible substance;
and (4): mixing micrococcus seeds and the residue mixed solution according to a volume-mass ratio of 1 (10-200), stirring at constant temperature, and fermenting for 12-96h to obtain a fermentation residue mixed solution, wherein the volume unit of the micrococcus seeds is mL, and the mass unit of the residue mixed solution is g;
and (5): filtering the mixed and dissolved substances of the fermentation residues to obtain a first filtrate and a first solid, and centrifuging the first filtrate to obtain a supernatant and a second solid;
and (6): mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to a mass-volume ratio of 1 (1-20), shaking at normal temperature for a period of time, and filtering to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL;
and (7): mixing the supernatant with the second filtrate to obtain a secondary tobacco extracting solution;
and (8): vacuum concentrating the secondary tobacco extract to density of 1.195-1.205g/mL to obtain a concentrated solution;
and (9): carrying out reflux extraction treatment on the concentrated solution to obtain a baked concentrated solution;
step (10): adding absolute ethyl alcohol into the baking concentrated solution, standing at low temperature for a period of time, centrifuging, and taking supernatant to obtain alcohol precipitate;
step (11): adding 1, 2-propylene glycol with the volume of 1-50% of the volume of the alcohol precipitate into the alcohol precipitate to obtain the tobacco secondary extract.
Optionally, the liquid culture medium in the step (1) is an LB liquid culture medium.
Optionally, the culturing conditions in step (1) are as follows:
the temperature is 30 ℃, the shaking speed is 150r/min, and the time is 12 h.
Optionally, the step (4) is specifically as follows:
mixing micrococcus seeds and residue miscible substances according to a volume-mass ratio of 1 (10-200), stirring and fermenting at a constant temperature of 30 ℃ for 12-96h at a stirring rotation speed of 180r/min to obtain fermentation residue miscible substances, wherein the volume unit of the micrococcus seeds is mL, and the mass unit of the residue miscible substances is g.
Optionally, the step (5) is specifically as follows:
step (5-1): filtering the mixed mixture of the fermentation residues to obtain a first filtrate and a first solid;
step (5-2): the first filtrate was centrifuged at 8000r/min for 20min at 4 ℃ to obtain a supernatant and a second solid.
Optionally, the step (6) is specifically as follows:
mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to the mass-volume ratio of 1 (1-20), shaking for 2h at the normal temperature at 150r/min, and filtering by using quick filter paper to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL.
Optionally, the step (8) is specifically as follows:
concentrating the secondary tobacco extractive solution at 50 deg.C and vacuum degree of 150mbar to density of 1.195-1.205g/mL to obtain concentrated solution.
Optionally, the step (9) is specifically as follows:
heating and baking the concentrated solution for 1-5h at 110 deg.C by reflux extraction device to obtain baked concentrated solution.
Optionally, the step (10) is specifically as follows:
adding 3 times of anhydrous ethanol into the roasted concentrated solution, standing overnight at-20 deg.C, centrifuging, and collecting supernatant to obtain ethanol precipitate.
Optionally, the step (11) is specifically as follows:
adding 1, 2-propylene glycol with volume of 10% of that of the alcohol precipitate to obtain tobacco secondary extract.
The secondary extraction method of the tobacco extract residues can effectively extract beneficial components in the tobacco extract residues to obtain the novel tobacco extract.
Other features of the present invention and advantages thereof will become apparent from the following detailed description of exemplary embodiments thereof, which proceeds with reference to the accompanying drawings.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description, serve to explain the principles of the invention.
FIG. 1 is a statistical chart of the flavor components of a tobacco secondary extract.
Detailed Description
Various exemplary embodiments of the present invention will now be described in detail with reference to the accompanying drawings. It should be noted that: the relative arrangement of the components and steps, the numerical expressions and numerical values set forth in these embodiments do not limit the scope of the present invention unless specifically stated otherwise.
The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses.
Techniques, methods, and apparatus known to those of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate.
In all examples shown and discussed herein, any particular value should be construed as merely illustrative, and not limiting. Thus, other examples of the exemplary embodiments may have different values.
The secondary extraction method of the tobacco extract residues provided by the disclosure comprises the following steps:
step (1): the micrococcus (Micrococcus Pribram) ZY02 with the preservation number of CGMCC NO.21313 is inoculated into a liquid culture medium and cultured for 6-36h in a shaker at 25-40 ℃ and 200 r/min.
The micrococcus (Microccceae Pribram) ZY02 is deposited in China general microbiological culture Collection center (CGMCC), the preservation number is CGMCC NO.21313, the strain name is Micrococcus, the strain number is ZY02, the classification name is Micrococcus, Microccceae Pribram, and the preservation time is 12 months and 07 days in 2020.
The liquid medium in step (1) may be an LB liquid medium.
The conditions for the culture in step (1) may be as follows:
the temperature is 30 ℃, the shaking speed is 150r/min, and the time is 12 h.
Step (2): centrifuging the seed solution at 4 deg.C at 4000r/min for 20min, removing supernatant, adding sterile water for redissolving, and adjusting OD 600 To 2.0, micrococcus seeds were prepared.
And (3): mixing the tobacco extract residue with deionized water according to the mass ratio of 1 (1-10) to obtain a residue miscible substance.
And (4): mixing micrococcus seeds and the residue miscible substance according to a volume-mass ratio of 1 (10-200), and fermenting for 12-96h under constant-temperature stirring to obtain a fermentation residue miscible substance, wherein the volume of the micrococcus seeds is mL, and the mass of the residue miscible substance is g.
The step (4) may be embodied as follows:
mixing micrococcus seeds and residue miscible substances according to a volume-mass ratio of 1 (10-200), stirring and fermenting at a constant temperature of 30 ℃ for 12-96h at a stirring rotation speed of 180r/min to obtain fermentation residue miscible substances, wherein the volume unit of the micrococcus seeds is mL, and the mass unit of the residue miscible substances is g.
And (5): filtering the mixed and dissolved substances of the fermentation residues to obtain a first filtrate and a first solid, and centrifuging the first filtrate to obtain a supernatant and a second solid.
The step (5) may be embodied as follows:
step (5-1): filtering the mixture of fermentation residues to obtain a first filtrate and a first solid.
Step (5-2): the first filtrate was centrifuged at 8000r/min for 20min at 4 ℃ to obtain a supernatant and a second solid.
And (6): and mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to the mass-to-volume ratio of 1 (1-20), shaking at normal temperature for a period of time, and filtering to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL.
The step (6) may be embodied as follows:
mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to the mass-volume ratio of 1 (1-20), shaking for 2h at the normal temperature at 150r/min, and filtering by using quick filter paper to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL.
And (7): and mixing the supernatant with the second filtrate to obtain a secondary tobacco extracting solution.
And (8): and (3) concentrating the secondary tobacco extracting solution in vacuum until the density is 1.195-1.205g/mL to obtain a concentrated solution.
The step (8) may be specifically as follows:
concentrating the secondary tobacco extractive solution at 50 deg.C and vacuum degree of 150mbar to density of 1.195-1.205g/mL to obtain concentrated solution.
And (9): and carrying out reflux extraction treatment on the concentrated solution to obtain a roasted concentrated solution.
The step (9) may be specifically as follows:
heating and baking the concentrated solution for 1-5h at 110 deg.C by reflux extraction device to obtain baked concentrated solution.
Step (10): adding absolute ethyl alcohol into the baking concentrated solution, standing at low temperature for a period of time, centrifuging, and taking supernatant to obtain alcohol precipitate.
The step (10) may be embodied as follows:
adding 3 times of anhydrous ethanol into the roasted concentrated solution, standing overnight at-20 deg.C, centrifuging, and collecting supernatant to obtain ethanol precipitate.
Step (11): adding 1, 2-propylene glycol with the volume of 1-50% of the alcohol precipitate into the alcohol precipitate to obtain the tobacco secondary extract.
The step (11) may be embodied as follows:
adding 1, 2-propylene glycol with volume of 10% of that of the alcohol precipitate into the alcohol precipitate to obtain the tobacco secondary extract.
The experimental procedures used in the examples below are conventional unless otherwise specified, the materials and reagents used therein are commercially available, and the equipment used in the experiments are well known to those skilled in the art without otherwise specified.
Example 1
A micrococcus (Micrococcus Pribram) ZY02 with the preservation number of CGMCC NO.21313 is inoculated into an LB liquid medium and cultured in a shaker at 30 ℃ and 150r/min for 12h to prepare a seed solution.
Centrifuging the seed solution at 4 deg.C at 4000r/min for 20min, removing supernatant, adding sterile water for redissolving, and adjusting OD 600 To 2.0, micrococcus seeds were prepared.
Mixing the tobacco extract residue with deionized water according to the mass ratio of 1:5 to obtain a residue miscible substance. The tobacco extract residue is solid tobacco residue generated after preparing tobacco extract from Yunnan Kunming 2019 CJ34 tobacco leaves by adopting an ethanol high-temperature reflux extraction method.
Mixing micrococcus seeds and residue mixed solution according to a volume-to-mass ratio (mL/g) of 1:20, and fermenting at constant temperature of 30 ℃ for 48 hours at a stirring speed of 180r/min to obtain fermentation residue mixed solution.
Filtering the mixture of fermentation residues to obtain a first filtrate and a first solid.
The first filtrate was centrifuged at 8000r/min for 20min at 4 ℃ to obtain a supernatant and a second solid.
Mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to a mass-to-volume ratio (g/mL) of 1:10, shaking for 2h at the normal temperature at 150r/min, and filtering by using quick filter paper to obtain a second filtrate.
Mixing the supernatant with the second filtrate to obtain the secondary tobacco extract.
Concentrating the secondary tobacco extractive solution at 50 deg.C and vacuum degree of 150mbar to density of 1.20 + -0.005 g/mL to obtain concentrated solution.
Heating and baking the concentrated solution for 1-5h at 110 deg.C by reflux extraction device to obtain baked concentrated solution.
Adding 3 times of anhydrous ethanol into the roasted concentrated solution, standing overnight at-20 deg.C, centrifuging, and collecting supernatant to obtain ethanol precipitate.
Adding 1, 2-propylene glycol with volume of 10% of that of the alcohol precipitate into the alcohol precipitate to obtain tobacco secondary extract T.
Comparative example 1
Mixing the tobacco extract residue with deionized water according to the mass ratio of 1:5 to obtain a residue miscible substance. The tobacco extract residue is solid tobacco powder residue generated after preparing tobacco extract from Yunnan Kunming 2019 CJ34 tobacco leaves by ethanol high-temperature reflux extraction method.
Mixing deionized water and the residue miscible substance according to a volume-to-mass ratio (mL/g) of 1:20, and stirring at a constant temperature of 30 ℃ for 48 hours at a stirring speed of 180r/min to obtain the residue miscible substance.
The residue mixture is filtered to obtain a first filtrate and a first solid.
Centrifuging the first filtrate at 8000r/min at 4 deg.C for 20min to obtain supernatant and second solid.
Mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to a mass-to-volume ratio (g/mL) of 1:10, shaking for 2h at the normal temperature at 150r/min, and filtering by using quick filter paper to obtain a second filtrate.
Mixing the supernatant with the second filtrate to obtain the secondary tobacco extract.
Concentrating the secondary tobacco extractive solution at 50 deg.C and vacuum degree of 150mbar to density of 1.20 + -0.005 g/mL to obtain concentrated solution.
Heating and baking the concentrated solution for 1-5h at 110 deg.C by reflux extraction device to obtain baked concentrated solution.
Adding 3 times of anhydrous ethanol into the roasted concentrated solution, standing overnight at-20 deg.C, centrifuging, and collecting supernatant to obtain ethanol precipitate.
Adding 1, 2-propylene glycol with the volume being 10% of the volume of the alcohol precipitate into the alcohol precipitate to obtain a tobacco secondary extract CK.
The conventional chemical components of the tobacco secondary extract in example 1 and comparative example 1 were measured by flow analysis, and the flavor components of the tobacco secondary extract T and the tobacco secondary extract CK were measured by GC-MS.
The conventional chemical components of the tobacco secondary extract are shown in table 1. As can be seen from Table 1, after fermentation, the contents of total sugar, reducing sugar, nicotine and total nitrogen in the secondary extract of tobacco are significantly increased, and the sugar-base ratio is increased. The results show that after fermentation, macromolecular substances are converted into soluble micromolecular sugar and amino acid, and the coated soluble substances are released along with the degradation of the macromolecules, so that the content in the tobacco secondary extract T is richer.
TABLE 1 analysis of the content of conventional chemical components of the secondary extract of tobacco (unit: mg/mL)
The flavor components of the tobacco secondary extract are shown in Table 2. As is clear from Table 2 and FIG. 1, the total content of flavor components of the fermented tobacco secondary extract T was higher than that of the unfermented tobacco secondary extract CK. The content of the Maillard product is increased most obviously, a certain amount of micromolecule saccharides and amino acids are generated by macromolecular degradation after fermentation, a large amount of Maillard products are generated by a high-temperature process, and the quality of the secondary extract of the tobacco is improved.
TABLE 2 statistics of the flavour content of the tobacco secondary extract (unit: ug/mL)
Although some specific embodiments of the present invention have been described in detail by way of examples, it should be understood by those skilled in the art that the above examples are for illustrative purposes only and are not intended to limit the scope of the present invention. It will be appreciated by those skilled in the art that modifications can be made to the above embodiments without departing from the scope and spirit of the invention. The scope of the invention is defined by the appended claims.
Claims (9)
1. A secondary extraction method of tobacco extract residues is characterized by comprising the following steps:
step (1): inoculating a colony of Micrococcus (Micrococcus epribram) ZY02 with the preservation number of CGMCC NO.21313 into a liquid culture medium, and culturing for 6-36h in a shaker at 25-40 ℃ and 200r/min of 100-;
step (2): centrifuging the seed solution at 4 deg.C at 4000r/min for 20min, removing supernatant, adding sterile water for redissolving, and adjusting OD 600 2.0, preparing micrococcus seeds;
and (3): mixing the tobacco extract residue with deionized water according to the mass ratio of 1 (1-10) to obtain a residue miscible substance;
and (4): mixing micrococcus seeds and the residue mixed solution according to a volume-mass ratio of 1 (10-200), stirring at constant temperature, and fermenting for 12-96h to obtain a fermentation residue mixed solution, wherein the volume unit of the micrococcus seeds is mL, and the mass unit of the residue mixed solution is g;
and (5): filtering the mixed and dissolved substances of the fermentation residues to obtain a first filtrate and a first solid, and centrifuging the first filtrate to obtain a supernatant and a second solid;
and (6): mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to the mass-to-volume ratio of 1 (1-20), shaking at normal temperature for a period of time, and filtering to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL;
and (7): mixing the supernatant with the second filtrate to obtain a secondary tobacco extracting solution;
and (8): vacuum concentrating the secondary tobacco extract until the density is 1.195-1.205g/mL to obtain a concentrated solution;
and (9): heating and baking the concentrated solution for 1-5h at 110 ℃ by adopting a reflux extraction device to obtain a baked concentrated solution;
step (10): adding absolute ethyl alcohol into the baking concentrated solution, standing at low temperature for a period of time, centrifuging, and taking supernatant to obtain alcohol precipitate;
step (11): adding 1, 2-propylene glycol with the volume of 1-50% of the alcohol precipitate into the alcohol precipitate to obtain the tobacco secondary extract.
2. The secondary extraction method of tobacco extract residue as claimed in claim 1, wherein the liquid culture medium in step (1) is LB liquid culture medium.
3. The method for secondary extraction of tobacco extract residue according to claim 1, wherein the conditions for culturing in the step (1) are as follows:
the temperature is 30 ℃, the oscillation speed is 150r/min, and the time is 12 h.
4. The method for secondary extraction of tobacco extract residue according to claim 1, wherein the step (4) is specifically as follows:
mixing micrococcus seeds and residue miscible substances according to a volume-mass ratio of 1 (10-200), stirring and fermenting at a constant temperature of 30 ℃ for 12-96h at a stirring rotation speed of 180r/min to obtain fermentation residue miscible substances, wherein the volume unit of the micrococcus seeds is mL, and the mass unit of the residue miscible substances is g.
5. The method for secondary extraction of tobacco extract residue according to claim 1, wherein the step (5) is specifically as follows:
step (5-1): filtering the mixture of the fermentation residues to obtain a first filtrate and a first solid;
step (5-2): the first filtrate was centrifuged at 8000r/min for 20min at 4 ℃ to obtain a supernatant and a second solid.
6. The method for secondary extraction of tobacco extract residue according to claim 1, wherein the step (6) is specifically as follows:
mixing the first solid and the second solid to obtain a third solid, mixing the third solid and absolute ethyl alcohol according to the mass-volume ratio of 1 (1-20), shaking for 2h at the normal temperature at 150r/min, and filtering by using quick filter paper to obtain a second filtrate, wherein the unit of the mass of the third solid is g, and the unit of the volume of the absolute ethyl alcohol is mL.
7. The secondary extraction method of tobacco extract residue as claimed in claim 1, wherein the step (8) is specifically as follows:
concentrating the secondary tobacco extractive solution at 50 deg.C under vacuum degree of 150mbar to density of 1.195-1.205g/mL to obtain concentrated solution.
8. The secondary extraction method of tobacco extract residue according to claim 1, wherein the step (10) is specifically as follows:
adding 3 times of anhydrous ethanol into the roasted concentrated solution, standing overnight at-20 deg.C, centrifuging, and collecting supernatant to obtain ethanol precipitate.
9. The method for secondary extraction of tobacco extract residue according to claim 1, wherein the step (11) is specifically as follows:
adding 1, 2-propylene glycol with volume of 10% of that of the alcohol precipitate into the alcohol precipitate to obtain the tobacco secondary extract.
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DE2816427C2 (en) * | 1977-05-06 | 1982-09-16 | Fabriques de Tabac Réunies S.A., 2003 Neuchâtel | Process for refining tobacco |
DE2811690C3 (en) * | 1977-05-06 | 1982-05-06 | Fabriques de Tabac Réunies S.A., 2003 Neuchâtel | Process for refining tobacco |
US4556073A (en) * | 1978-06-15 | 1985-12-03 | Brown & Williamson Tobacco Corporation | Process for reduction of nitrate content of tobacco by microbial treatment |
US4709710A (en) * | 1978-09-05 | 1987-12-01 | Fabriques De Tabac Reunies S.A. | Process for improving tobacco |
US4572219A (en) * | 1979-01-19 | 1986-02-25 | Fabriques De Tabac Reunies S.A. | Process for improving tobacco |
CA1140876A (en) * | 1979-06-15 | 1983-02-08 | Lawrence E. Gravely | Process for reduction of nitrate content of tobacco by microbial treatment |
CN101449854B (en) * | 2008-12-30 | 2011-12-21 | 无锡华馨香精香料有限公司 | Fermentation optimization method of tobacco essence |
CN109797114B (en) * | 2017-11-17 | 2022-04-29 | 中国科学院青岛生物能源与过程研究所 | Micrococcus and application thereof in production of 2, 3-butanediol |
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